Hong Kong Science and Technology Park 1 article published in JoVE Biology Establishing Human Lung Organoids and Proximal Differentiation to Generate Mature Airway Organoids Cun Li1,2, Man Chun Chiu1,2, Yifei Yu1,2, Xiaojuan Liu1,2, Ding Xiao1,2, Jingjing Huang1,2, Zhixin Wan1,2, Jie Zhou1,2,3 1Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, 2Centre for Virology, Vaccinology and Therapeutics, Hong Kong Science and Technology Park, 3State Key Laboratory of Emerging Infectious Diseases, The University of Hong Kong The protocol presents a method to derive human lung organoids from primary lung tissues, expand the lung organoids and induce proximal differentiation to generate 3D and 2D airway organoids that faithfully phenocopy the human airway epithelium.